Crystalline forms of an androgen receptor modulator

ABSTRACT

Described herein are amorphous and crystalline forms of the androgen receptor modulator 4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide. Also described are pharmaceutical compositions suitable for administration to a mammal that include the androgen receptor modulator, and methods of using the androgen receptor modulator, alone and in combination with other compounds, for treating diseases or conditions that are associated with androgen receptor activity.

RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 16/507,780, filed Jul. 10, 2019, which is divisionalapplication of U.S. patent application Ser. No. 16/384,002, filed Apr.15, 2019, which is a divisional application of U.S. patent applicationSer. No. 15/975,449, filed May 9, 2018, now U.S. Pat. No. 10,308,630that issued Jun. 4, 2019, which is a divisional application of U.S.patent application Ser. No. 15/262,522, filed Sep. 12, 2016, now U.S.Pat. No. 9,994,545 that issued on Jun. 12, 2018, which is a divisionalapplication of U.S. patent application Ser. No. 14/406,520, filed Dec.8, 2014, now U.S. Pat. No. 9,481,663 that issued Nov. 1, 2016, which isa U.S. National Stage Application of PCT/US2013/044116, filed Jun. 4,2013, which claims the benefit of U.S. Provisional Patent ApplicationSer. No. 61/656,888, filed on Jun. 7, 2012, each of which is hereinincorporated by reference in its entirety.

FIELD OF THE INVENTION

Described herein are crystalline forms of the androgen receptormodulator4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,pharmaceutically acceptable salts, solvates, as well as pharmaceuticalcompositions thereof, and methods of use thereof in the treatment orprevention of diseases or conditions associated with androgen receptoractivity.

BACKGROUND OF THE INVENTION

The androgen receptor (“AR”) is a ligand-activated transcriptionalregulatory protein that mediates induction of a variety of biologicaleffects through its interaction with endogenous androgens. Endogenousandrogens include steroids such as testosterone and dihydrotestosterone.Testosterone is converted to dihydrotestosterone by the enzyme 5alpha-reductase in many tissues.

The actions of androgens with androgen receptors have been implicated ina number of diseases or conditions, such as androgen dependent cancers,virilization in women, and acne, among others. Compounds that diminishthe effects of androgens with androgen receptors and/or lower theconcentrations of androgen receptors find use in the treatment ofdiseases or conditions in which androgen receptors play a role.

SUMMARY OF THE INVENTION

Described herein is4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or a pharmaceutically acceptable salt thereof, including allpharmaceutically acceptable solvates (including hydrates), polymorphs,and amorphous phases thereof, and methods of uses thereof4-[7-(6-Cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,as well as pharmaceutically acceptable salts thereof, is used in themanufacture of medicaments for the treatment or prevention of diseases,disorders, or conditions associated with androgen receptor activity.

Described herein are pharmaceutical compositions comprising4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or a pharmaceutically acceptable salt thereof as the active ingredientin the pharmaceutical composition.

In one aspect, described herein is crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form A. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form B. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form C. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form D. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form E. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form F. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form G. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form H. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form I. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form J.

In some embodiments, described herein is a pharmaceutically acceptablesalt of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,wherein the pharmaceutically acceptable salt is an acid addition salt.In some embodiments, the pharmaceutically acceptable salt is amorphous.In some embodiments, the pharmaceutically acceptable salt iscrystalline.

In some embodiments, described herein is a pharmaceutical compositioncomprising a crystalline form of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideas described herein, and at least one additional ingredient selectedfrom pharmaceutically acceptable carriers, diluents and excipients. Insome embodiments, the pharmaceutical composition includes Form A of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In some embodiments, the pharmaceutical composition includes Form B. Insome embodiments, the pharmaceutical composition includes Form C. Insome embodiments, the pharmaceutical composition includes Form D. Insome embodiments, the pharmaceutical composition includes Form E. Insome embodiments, the pharmaceutical composition includes Form F. Insome embodiments, the pharmaceutical composition includes Form G. Insome embodiments, the pharmaceutical compositions includes Form H. Insome embodiments, the pharmaceutical composition includes Form I. Insome embodiments, the pharmaceutical composition includes Form J. Insome embodiments, the pharmaceutical composition is in a form suitablefor oral administration to a mammal. In some embodiments, thepharmaceutical composition is in an oral dosage form. In someembodiments, the pharmaceutical composition is in an oral solid dosageform. In some embodiments, the pharmaceutical composition is in the formof a tablet, pill, or capsule. In some embodiments, the pharmaceuticalcomposition is in the form of a capsule. In some embodiments, thepharmaceutical composition is in the form of an immediate releasecapsule or an enteric coated capsule. In some embodiments, thepharmaceutical composition is in the form of a tablet. In someembodiments, the pharmaceutical composition is in the form of animmediate release tablet, an enteric coated tablet, or a sustainedrelease tablet. In some embodiments, the pharmaceutical composition isin the form of a moisture barrier coated tablet. In some embodiments,the pharmaceutical composition comprises about 0.5 mg to about 1000 mgof crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In some embodiments, the pharmaceutical composition comprises about 30mg to about 300 mg of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.

Also provided is an article of manufacture comprising multiple unitdoses of the oral solid dosage form pharmaceutical composition describedherein in a high-density polyethylene (HDPE) bottle equipped with ahigh-density polyethylene (HDPE) cap. In some embodiments, high-densitypolyethylene (HDPE) bottle further comprises an aluminum foil inductionseal and silica gel desiccant.

Also described is a method of treating prostate cancer in a mammalcomprising administering to the mammal a pharmaceutical composition asdescribed herein. In some embodiments, the prostate cancer is hormonesensitive prostate cancer or hormone refractory prostate cancer.

Also provided is the use of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or a pharmaceutically acceptable salt thereof, for the manufacture of amedicament for the treatment or prevention of prostate cancer in ahuman. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline.

Also described herein are processes for the preparation of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.The disclosed processes provide for the preparation of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein good yield and high purity.

Other objects, features and advantages of the methods and compositionsdescribed herein will become apparent from the following detaileddescription. It should be understood, however, that the detaileddescription and the specific examples, while indicating specificembodiments, are given by way of illustration only, since variouschanges and modifications within the spirit and scope of the inventionwill become apparent to those skilled in the art from this detaileddescription.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 illustrates the XRPD of Form A.

FIG. 2 illustrates the XRPD of Form B.

FIG. 3 illustrates the XRPD of Form C.

FIG. 4 illustrates the XRPD of Form D.

FIG. 5 illustrates the XRPD of Form E.

FIG. 6 illustrates the XRPD of Form F.

FIG. 7 illustrates the XRPD of Form G.

FIG. 8 illustrates the XRPD of Form H.

FIG. 9 illustrates the XRPD of Form I.

FIG. 10 illustrates the XRPD of Form J.

FIG. 11 illustrates the TGA and DSC thermograms of Form B.

FIG. 12 illustrates the TGA and DSC thermograms of Form C.

FIG. 13 illustrates the TGA and DSC thermograms of Form D.

FIG. 14 illustrates the TGA and DSC thermograms of Form E.

FIG. 15 illustrates the TGA and DSC thermograms of Form F.

FIG. 16 illustrates the DSC thermogram of Form G.

FIG. 17 illustrates the TGA and DSC thermograms of Form H.

FIG. 18 illustrates the TGA and DSC thermograms of Form J.

FIG. 19 illustrates the DSC thermogram of Form A.

DETAILED DESCRIPTION OF THE INVENTION

The androgen receptor (AR) is a member of the nuclear receptorsuperfamily. Among this family of proteins, only five vertebrate steroidreceptors are known and include the androgen receptor, estrogenreceptor, progesterone receptor, glucocorticoid receptor, andmineralocorticoid receptor. AR is a soluble protein that functions as anintracellular transcriptional factor. AR function is regulated by thebinding of androgens, which initiates sequential conformational changesof the receptor that affect receptor-protein interactions andreceptor-DNA interactions.

AR is mainly expressed in androgen target tissues, such as the prostate,skeletal muscle, liver, and central nervous system (CNS), with higherexpression levels observed in the prostate, adrenal gland, andepididymis. AR can be activated by the binding of endogenous androgens,including testosterone and 5α-dihydrotestosterone (5α-DHT).

The androgen receptor (AR), located on Xq11-12, is a 110 kD nuclearreceptor that, upon activation by androgens, mediates transcription oftarget genes that modulate growth and differentiation of prostateepithelial cells. Similar to the other steroid receptors, unbound AR ismainly located in the cytoplasm and associated with a complex of heatshock proteins (HSPs) through interactions with the ligand-bindingdomain. Upon agonist binding, AR goes through a series of conformationalchanges: the heat shock proteins dissociate from AR, and the transformedAR undergoes dimerization, phosphorylation, and translocation to thenucleus, which is mediated by the nuclear localization signal.Translocated receptor then binds to the androgen response element (ARE),which is characterized by the six-nucleotide half-site consensussequence 5′-TGTTCT-3′ spaced by three random nucleotides and is locatedin the promoter or enhancer region of AR gene targets. Recruitment ofother transcription co-regulators (including co-activators andco-repressors) and transcriptional machinery further ensures thetransactivation of AR-regulated gene expression. All of these processesare initiated by the ligand-induced conformational changes in theligand-binding domain.

AR signaling is crucial for the development and maintenance of malereproductive organs including the prostate gland, as genetic malesharboring loss of function AR mutations and mice engineered with ARdefects do not develop prostates or prostate cancer. This dependence ofprostate cells on AR signaling continues even upon neoplastictransformation. Androgen depletion (using GnRH agonists) continues to bethe mainstay of prostate cancer treatment. However androgen depletion isusually effective for a limited duration and prostate cancer evolves toregain the ability to grow despite low levels of circulating androgens.Treatment options for castration resistant prostate cancer (CRPC) arelimited, with docetaxel and abiraterone acetate (a CYP17 inhibitor)being agents that have been shown to prolong survival. Interestingly,while a small minority of CRPC does bypass the requirement for ARsignaling, the vast majority of CRPC, though frequently termed “androgenindependent prostate cancer” or “hormone refractory prostate cancer,”retains its lineage dependence on AR signaling.

Prostate cancer is the second most common cause of cancer death in menin the US, and approximately one in every six American men will bediagnosed with the disease during his lifetime. Treatment aimed ateradicating the tumor is unsuccessful in 30% of men, who developrecurrent disease that is usually manifest first as a rise in plasmaprostate-specific antigen (PSA) followed by spread to distant sites.Given that prostate cancer cells depend on androgen receptor (AR) fortheir proliferation and survival, these men are treated with agents thatblock production of testosterone (e.g. GnRH agonists), alone or incombination with anti-androgens (e.g. bicalutamide), which antagonizethe effect of any residual testosterone. The approach is effective asevidenced by a drop in PSA and regression of visible tumor (if present);however, this is followed by regrowth as a “castration resistant”prostate cancer (CRPC) to which most patients eventually succumb. Recentstudies on the molecular basis of CRPC have demonstrated that CRPCcontinues to depend on AR signaling and that a key mechanism of acquiredresistance is an elevated level of AR protein (Nat. Med, 2004, 10,33-39). AR targeting agents with activity in hormone sensitive andcastration resistant prostate cancer have great promise in treating thislethal disease.

Anti-androgens are useful for the treatment of prostate cancer duringits early stages. However, prostate cancer often advances to ahormone-refractory state in which the disease progresses in the presenceof continued androgen ablation or anti-androgen therapy. Instances ofanti-androgen withdrawal syndrome have also been reported afterprolonged treatment with anti-androgens. Anti-androgen withdrawalsyndrome is commonly observed clinically and is defined in terms of thetumor regression or symptomatic relief observed upon cessation ofanti-androgen therapy. AR mutations that result in receptor promiscuityand the ability of these anti-androgens to exhibit agonist activitymight at least partially account for this phenomenon. For example,hydroxyflutamide and bicalutamide act as AR agonists in T877A andW741L/W741C AR mutants, respectively.

In the setting of prostate cancer cells that were rendered “castrationresistant” via over expression of AR, it has been demonstrated thatcertain anti-androgen compounds, such as bicalutamide, have noantagonist activity, but instead have modest agonist activity (Science,2009 May 8; 324(5928): 787-790). This agonist activity helps to explaina clinical observation, called the anti-androgen withdrawal syndrome,whereby about 30% of men who progress on AR antagonists experience adecrease in serum PSA when therapy is discontinued (J Clin Oncol, 1993.11(8): p. 1566-72).

Given the central role of AR in prostate cancer development andprogression,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis useful in the treatment of prostate cancer.

AR-related diseases or conditions include, but are not limited to,benign prostate hyperplasia, hirsutism, acne, adenomas and neoplasias ofthe prostate, benign or malignant tumor cells containing the androgenreceptor, hyperpilosity, seborrhea, endometriosis, polycystic ovarysyndrome, androgenic alopecia, hypogonadism, osteoporosis, suppressionof spermatogenesis, libido, cachexia, anorexia, androgen supplementationfor age related decreased testosterone levels, prostate cancer, breastcancer, endometrial cancer, uterine cancer, hot flashes, Kennedy'sdisease muscle atrophy and weakness, skin atrophy, bone loss, anemia,arteriosclerosis, cardiovascular disease, loss of energy, loss ofwell-being, type 2 diabetes, and abdominal fat accumulation.

4-[7-(6-Cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis an androgen receptor modulator that inhibits both AR nucleartranslocation and AR binding to androgen response elements in DNA.Importantly, and in contrast to the first-generation anti-androgenbicalutamide, it exhibits no agonist activity in prostate cancer cellsthat over-express androgen receptors. It is well suited as either amono- or a combination therapy across the entire spectrum of prostatecancer disease states.

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis used to treat prostate cancer in a mammal, wherein the mammal ischemotherapy-nave.

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis used to treat prostate cancer in a mammal, wherein the mammal isbeing treated for prostate cancer with at least one anti-cancer agent.In one embodiment, the prostate cancer is hormone refractory prostatecancer. In one embodiment, the prostate cancer is bicalutamide-resistantprostate cancer.

4[7-(6-Cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,and Pharmaceutically Acceptable Salts Thereof

“4-[7-(6-Cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide”refers to the compound with the following structure:

A wide variety of pharmaceutically acceptable salts of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideare possible and include acid addition salts, that are formed byreacting the free base of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidewith an inorganic acid or an organic acid. Such salt forms of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideinclude, but are not limited to: hydrochloric acid salt, hydrobromicacid salt, sulfuric acid salt, phosphoric acid salt, metaphosphoric acidsalt, acetic acid salt, propionic acid salt, hexanoic acid salt,cyclopentanepropionic acid salt, glycolic acid salt, pyruvic acid salt,lactic acid salt, malonic acid salt, succinic acid salt, malic acidsalt, maleic acid salt, fumaric acid salt, trifluoroacetic acid salt,tartaric acid salt, citric acid salt, benzoic acid salt,3-(4-hydroxybenzoyl)benzoic acid salt, cinnamic acid salt, mandelic acidsalt, methanesulfonic acid salt, ethanesulfonic acid salt,1,2-ethanedisulfonic acid salt, 2-hydroxyethanesulfonic acid salt,benzenesulfonic acid salt, toluenesulfonic acid salt,2-naphthalenesulfonic acid salt,4-methylbicyclo-[2.2.2]oct-2-ene-1-carboxylic acid salt, glucoheptonicacid salt, 3-phenylpropionic acid salt, trimethylacetic acid salt,tertiary butylacetic acid salt, lauryl sulfuric acid salt, gluconic acidsalt, glutamic acid salt, hydroxynaphthoic acid salt, salicylic acidsalt, stearic acid salt, muconic acid salt, butyric acid salt,phenylacetic acid salt, phenylbutyric acid salt, valproic acid salt, andthe like.

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis used in any of the pharmaceutical compositions or methods describedherein.

In some embodiments, a pharmaceutically acceptable salt of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis used in any of the pharmaceutical compositions or methods describedherein.

The term “pharmaceutically acceptable salt” in reference to4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamiderefers to a salt of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,which does not cause significant irritation to a mammal to which it isadministered and does not substantially abrogate the biological activityand properties of the compound.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms (solvates). Solvatescontain either stoichiometric or non-stoichiometric amounts of asolvent, and are formed during the process of product formation orisolation with pharmaceutically acceptable solvents such as water,ethanol, methyl tert-butyl ether, isopropanol, acetonitrile, heptane,and the like. In one aspect, solvates are formed using, but not limitedto, Class 3 solvent(s). Categories of solvents are defined in, forexample, the International Conference on Harmonization of TechnicalRequirements for Registration of Pharmaceuticals for Human Use (ICH),“Impurities: Guidelines for Residual Solvents, Q3C(R3), (November 2005).Hydrates are formed when the solvent is water, or alcoholates are formedwhen the solvent is an alcohol. In one embodiment, solvates of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or salts thereof, are conveniently prepared or formed during theprocesses described herein. In other embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or salts thereof, exist in unsolvated form.

In yet other embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideor a pharmaceutically acceptable salt thereof is prepared in variousforms, including but not limited to, amorphous phase, milled forms, andnano-particulate forms.

Amorphous4-[7-(6-Cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis amorphous. In some embodiments, Amorphous Phase of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidehas an XRPD pattern showing a lack of crystallinity.

Form A

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form A. Form A of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as exhibiting at least one of:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 1;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 4.8±0.1° 2-Theta, 7.1±0.1° 2-Theta, 14.2±0.1° 2-Theta,    16.3±0.1° 2-Theta, 20.1±0.1° 2-Theta;-   (c) unit cell parameters substantially equal to the following at    −173° C.:

Crystal system Orthorhombic Space group P2(1)2(1)2 a 16.3429(3) Å α 90°b 37.7298(7) Å β 90° c 7.23410(10) Å γ 90° V 4460.65(13) Å3 Z 8 Dc 1.446g · cm⁻¹

-   (d) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;-   (e) a DSC thermogram with an endotherm having an onset temperature    at about 108-120° C. and a peak at about 133-135° C.;-   (f) a DSC thermogram substantially similar to the one set forth in    FIG. 19;-   (g) an observed aqueous solubility of about 0.01 mg/mL;    -   or-   (h) combinations thereof.

In some embodiments, Form A is characterized as exhibiting at least two,at least three, at least four, at least five, at least six or all sevenof the properties selected from (a) to (g). In some embodiments, Form Ais characterized as having properties (a), (b), (c), (d), (e), (f) and(g). In some embodiments, Form A is characterized as having property(a), (b), (c), (d), (g) or combinations thereof. In some embodiments,Form A is characterized as having at least two, at least three, at leastfour or all five of the properties selected from (a), (b), (c), (d), and(g). In some embodiments, Form A is characterized as having properties(a), (b), (c), (d), and (g).

In some embodiments, Form A is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 1. Insome embodiments, Form A is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 4.8±0.1°2-Theta, 7.1±0.1° 2-Theta, 14.2±0.1° 2-Theta, 16.3±0.1° 2-Theta,20.1±0.1° 2-Theta. In some embodiments, Form A is characterized ashaving substantially the same X-ray powder diffraction (XRPD) patternpost storage at 40° C. and 75% RH for at least a week.

In some embodiments, Form A is characterized as having unit cellparameters substantially equal to the following at −173° C.:

Crystal system Orthorhombic Space group P2(1)2(1)2 a 16.3429(3) Å α 90°b 37.7298(7) Å β 90° c 7.23410(10) Å γ 90° V 4460.65(13) Å3 Z 8 Dc 1.446g · cm⁻¹

In some embodiments, Form A is characterized as having a DSC thermogramwith an endotherm having an onset temperature at about 108-120° C. and apeak at about 133-135° C.;

In some embodiments, Form A is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 19.

In some embodiments, Form A is characterized as having an observedaqueous solubility of about 0.01 mg/mL

In some embodiments, Form A is obtained from ethanol, tetrahydrofuran(THF), dichloromethane, acetone, methanol, nitromethane, water,THF-water mixture, or dioxane-water mixture. In some embodiments, Form Ais obtained from ethanol. In some embodiments, Form A is solvated. Insome embodiments, Form A is an ethanol solvate. In some embodiments,Form A is unsolvated. In some embodiments, Form A is a hydrate. In someembodiments, Form A is a solvated hydrate.

Form B

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form B. Form B is unsolvated. Form B of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 2;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 12.1±0.1° 2-Theta, 16.0±0.1° 2-Theta, 16.7±0.1° 2-Theta,    20.1±0.1° 2-Theta, 20.3±0.1° 2-Theta;-   (c) unit cell parameters substantially equal to the following at    −173° C.:

Crystal system Monoclinic Space group P2₁/c a 17.7796(4) Å α 90° b12.9832(3) Å β 100.897(2)° c 18.4740(4) Å γ 90° V 4187.57(16) Å³ Z 8 Dc1.515 g · cm⁻¹

-   (d) a DSC thermogram substantially similar to the one set forth in    FIG. 11;-   (e) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 11;-   (f) a DSC thermogram with an endotherm having an onset temperature    at about 194° C.;-   (g) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;-   (h) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 25° C. and 92% RH for 12 days;-   (i) an observed aqueous solubility of about 0.004 mg/mL;-    or-   (j) combinations thereof.

In some embodiments, Form B is characterized as having at least two, atleast three, at least four, at least five, at least six, at least seven,at least eight or all nine of the properties selected from (a) to (i).

In some embodiments, Form B is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 2. Insome embodiments, Form B is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 12.1±0.1°2-Theta, 16.0±0.1° 2-Theta, 16.7±0.1° 2-Theta, 20.1±0.1° 2-Theta,20.3±0.1° 2-Theta. In some embodiments, Form B is characterized ashaving substantially the same X-ray powder diffraction (XRPD) patternpost storage at 40° C. and 75% RH for at least a week. In someembodiments, Form B is characterized as having substantially the sameX-ray powder diffraction (XRPD) pattern post storage at 25° C. and 92%RH for 12 days.

In some embodiments, Form B is characterized as having unit cellparameters substantially equal to the following at −173° C.:

Crystal system Monoclinic Space group P2₁/c a 17.7796(4) Å α 90° b12.9832(3) Å β 100.897(2)° c 18.4740(4) Å γ 90° V 4187.57(16) Å³ Z 8 Dc1.515 g · cm⁻¹

In some embodiments, Form B is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 11. In someembodiments, Form B is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 11. In some embodiments, Form B is characterized as having a DSCthermogram with an endotherm having an onset temperature at about 194°C.

In some embodiments, Form B is characterized as having an observedaqueous solubility of about 0.004 mg/mL.

In some embodiments, Form B is obtained from water, ethyl acetate,tert-butyl methyl ether (TBME), toluene, isopropylacetate, or methylethyl ketone (MEK).

Form C

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form C. Form C of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 3;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 4.3±0.1° 2-Theta, 6.9±0.1° 2-Theta, 9.1±0.1° 2-Theta,    10.6±0.1° 2-Theta, 13.8±0.1° 2-Theta, 26.4±0.1° 2-Theta;-   (c) a DSC thermogram substantially similar to the one set forth in    FIG. 12;-   (d) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 12;-   (e) a DSC thermogram with a first endotherm having an onset    temperature at about 118° C. and second endotherm having an onset    temperature at about 193° C.;-   (f) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;    or-   (g) combinations thereof.

In some embodiments, Form C is characterized as having at least two, atleast three, at least four, at least five, or all six of the propertiesselected from (a) to (f).

In some embodiments, Form C is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 3. Insome embodiments, Form C is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 4.3±0.1°2-Theta, 6.9±0.1° 2-Theta, 9.1±0.1° 2-Theta, 10.6±0.1° 2-Theta,13.8±0.1° 2-Theta, 26.4±0.1° 2-Theta. In some embodiments, Form C ischaracterized as having substantially the same X-ray powder diffraction(XRPD) pattern post storage at 40° C. and 75% RH for at least a week.

In some embodiments, Form C is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 12. In someembodiments, Form C is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 12. In some embodiments, Form C is characterized as having a DSCthermogram with a first endotherm having an onset temperature at about118° C. and second endotherm having an onset temperature at about 193°C.

In some embodiments, Form C is obtained from isopropanol (IPA), anisole,or IPA-water mixture. In some embodiments, Form C is solvated. In someembodiments, Form C is an isopropanol solvate.

Form D

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form D. Form D of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 4;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 6.3±0.1° 2-Theta, 13.9±0.1° 2-Theta, 16.4±0.1° 2-Theta,    17.0±0.1° 2-Theta, 23.7±0.1° 2-Theta, 24.8±0.1° 2-Theta;-   (c) a DSC thermogram substantially similar to the one set forth in    FIG. 13;-   (d) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 13;-   (e) a DSC thermogram with a first endotherm having an onset    temperature at about 122° C. and second endotherm having an onset    temperature at about 192° C.;-   (f) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;    or-   (g) combinations thereof.

In some embodiments, Form D is characterized as having at least two, atleast three, at least four, at least five, or all six of the propertiesselected from (a) to (f).

In some embodiments, Form D is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 4. Insome embodiments, Form D is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 6.3±0.1°2-Theta, 13.9±0.1° 2-Theta, 16.4±0.1° 2-Theta, 17.0±0.1° 2-Theta,23.7±0.1° 2-Theta, 24.8±0.1° 2-Theta. In some embodiments, Form D ischaracterized as having substantially the same X-ray powder diffraction(XRPD) pattern post storage at 40° C. and 75% RH for at least a week.

In some embodiments, Form D is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 13. In someembodiments, Form D is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 13. In some embodiments, Form D is characterized as having a DSCthermogram with an endotherm having an onset temperature at about 122°C. In some embodiments, Form D is characterized as having a DSCthermogram with a first endotherm having an onset temperature at about122° C. and second endotherm having an onset temperature at about 192°C.

In some embodiments, Form D is obtained from tert-butyl methyl ether(TBME). In some embodiments, Form D is solvated. In some embodiments,Form D is a tert-butyl methyl ether (TBME) solvate.

Form E

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form E. Form E of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 5;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 7.2±0.1° 2-Theta, 11.8±0.1° 2-Theta, 16.1±0.1° 2-Theta,    20.5±0.1° 2-Theta, 23.0±0.1° 2-Theta, 25.2±0.1° 2-Theta;-   (c) unit cell parameters substantially equal to the following at    −173° C.:

Crystal system Orthorhombic Space group P_(na)2₁ a 8.43080(10) Å  α 90°b 17.1685(3) Å β 90° c 17.4276(3) Å γ 90° V 2522.54(7) Å³ Z 4 Dc 1.463 g· cm⁻¹

-   (d) a DSC thermogram substantially similar to the one set forth in    FIG. 14;-   (e) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 14;-   (f) a DSC thermogram with an endotherm having an onset temperature    at about 116° C.;    or-   (g) combinations thereof.

In some embodiments, Form E is characterized as having at least two, atleast three, at least four, at least five, or all six of the propertiesselected from (a) to (f).

In some embodiments, Form E is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 5. Insome embodiments, Form E is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 7.2±0.1°2-Theta, 11.8±0.1° 2-Theta, 16.1±0.1° 2-Theta, 20.5±0.1° 2-Theta,23.0±0.1° 2-Theta, 25.2±0.1° 2-Theta.

In some embodiments, Form E is characterized as having unit cellparameters substantially equal to the following at −173° C.:

Crystal system Orthorhombic Space group P_(na)2₁ a 8.43080(10) Å  α 90°b 17.1685(3) Å β 90° c 17.4276(3) Å γ 90° V 2522.54(7) Å³ Z 4 Dc 1.463 g· cm⁻¹

In some embodiments, Form E is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 14. In someembodiments, Form E is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 14. In some embodiments, Form E is characterized as having a DSCthermogram with an endotherm having an onset temperature at about 116°C.

In some embodiments, Form E is obtained from dimethylsulfoxide. In someembodiments, Form E is solvated. In some embodiments, Form E is adimethylsulfoxide solvate.

Form F

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form F. Form F of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 6;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 4.6±0.1° 2-Theta, 6.1±0.1° 2-Theta, 14.3±0.1° 2-Theta,    21.6±0.1° 2-Theta, 22.4±0.1° 2-Theta, 23.3±0.1° 2-Theta, 25.5±0.1°    2-Theta;-   (c) a DSC thermogram substantially similar to the one set forth in    FIG. 15;-   (d) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 15;-   (e) a DSC thermogram with an endotherm having an onset temperature    at about 113° C.;    or-   (f) combinations thereof.

In some embodiments, Form F is characterized as having at least two, atleast three, at least four, or all five of the properties selected from(a) to (e).

In some embodiments, Form F is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 6. Insome embodiments, Form F is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 4.6±0.1°2-Theta, 6.1±0.1° 2-Theta, 14.3±0.1° 2-Theta, 21.6±0.1° 2-Theta,22.4±0.1° 2-Theta, 23.3±0.1° 2-Theta, 25.5±0.1° 2-Theta.

In some embodiments, Form F is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 15. In someembodiments, Form F is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 15. In some embodiments, Form F is characterized as having a DSCthermogram with an endotherm having an onset temperature at about 113°C.

In some embodiments, Form F is obtained from an acetone/water mixture.

Form G

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form G. Form G of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 7;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 7.0±0.1° 2-Theta, 10.3±0.1° 2-Theta, 14.1±0.1° 2-Theta,    15.2±0.1° 2-Theta, 23.6±0.1° 2-Theta;-   (c) unit cell parameters substantially equal to the following at    −173° C.:

Crystal system Monoclinic Space group Cc a 18.613(2) Å  α 90° b16.9728(14) Å  β 91.328(8)° c 7.8214(7) Å, γ 90° V 2470.2(4) Å³ Z 4 Dc1.488 g · cm⁻¹

-   (d) a DSC thermogram substantially similar to the one set forth in    FIG. 16;-   (e) a DSC thermogram with a first endotherm having an onset    temperature at about 101° C. and second endotherm having an onset    temperature at about 190° C.;-   (f) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;    or-   (g) combinations thereof.

In some embodiments, Form G is characterized as having at least two, atleast three, at least four, at least five, or all six of the propertiesselected from (a) to (f).

In some embodiments, Form G is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 7. Insome embodiments, Form G is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 7.0±0.1°2-Theta, 10.3±0.1° 2-Theta, 14.1±0.1° 2-Theta, 15.2±0.1° 2-Theta,23.6±0.1° 2-Theta.

In some embodiments, Form G is characterized as having unit cellparameters substantially equal to the following at −173° C.:

Crystal system Monoclinic Space group Cc a 18.613(2) Å α 90° b16.9728(14) Å  β 91.328(8)° c 7.8214(7) Å, γ 90° V 2470.2(4) Å³ Z 4 Dc1.488 g · cm⁻¹

In some embodiments, Form G is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 16. In someembodiments, Form G is characterized as having a DSC thermogram with anendotherm having an onset temperature at about 101° C. In someembodiments, Form G is characterized as having a DSC thermogram with afirst endotherm having an onset temperature at about 101° C. and secondendotherm having an onset temperature at about 190° C.

In some embodiments, Form G is characterized as having substantially thesame X-ray powder diffraction (XRPD) pattern post storage at 40° C. and75% RH for at least a week.

In some embodiments, Form G is obtained from 2-methoxyethanol. In someembodiments, Form G is solvated. In some embodiments, Form G is a2-methoxyethanol solvate.

Form H

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form H. Form H is unsolvated. Form H of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 8;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 8.0±0.1° 2-Theta, 14.7±0.1° 2-Theta, 15.9±0.1° 2-Theta,    18.2±0.1° 2-Theta, 25.7±0.1° 2-Theta, 26.7±0.1° 2-Theta;-   (c) a DSC thermogram substantially similar to the one set forth in    FIG. 17;-   (d) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 17;-   (e) a DSC thermogram with a first endotherm having an onset    temperature at about 173° C. and second endotherm having an onset    temperature at about 193° C.;-   (f) substantially the same X-ray powder diffraction (XRPD) pattern    post storage at 40° C. and 75% RH for at least a week;    or-   (g) combinations thereof.

In some embodiments, Form H is characterized as having at least two, atleast three, at least four, at least five, or all six of the propertiesselected from (a) to (f).

In some embodiments, Form H is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 8. Insome embodiments, Form H is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 8.0±0.1°2-Theta, 14.7±0.1° 2-Theta, 15.9±0.1° 2-Theta, 18.2±0.1° 2-Theta,25.7±0.1° 2-Theta, 26.7±0.1° 2-Theta. In some embodiments, Form H ischaracterized as having substantially the same X-ray powder diffraction(XRPD) pattern post storage at 40° C. and 75% RH for at least a week.

In some embodiments, Form H is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 17. In someembodiments, Form H is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 17. In some embodiments, Form H is characterized as having a DSCthermogram with a first endotherm having an onset temperature at about173° C. and second endotherm having an onset temperature at about 193°C.

In some embodiments, Form H is obtained from ethyl acetate.

Form I

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form I. Form I of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 9;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks 7.7±0.1° 2-Theta, 10.4±0.1° 2-Theta, 11.6±0.1° 2-Theta,    17.0±0.1° 2-Theta, 20.0±0.1° 2-Theta, 20.6±0.1° 2-Theta;    or-   (c) combinations thereof.

In some embodiments, Form I is characterized as having at least property(a) and property (b).

In some embodiments, Form I is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 9. Insome embodiments, Form I is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks 7.7±0.1° 2-Theta,10.4±0.1° 2-Theta, 11.6±0.1° 2-Theta, 17.0±0.1° 2-Theta, 20.0±0.1°2-Theta, 20.6±0.1° 2-Theta.

In some embodiments, Form I is obtained from dimethylsulfoxide.

Form J

In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis crystalline. In some embodiments,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis Form J. Form J of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis characterized as having:

-   (a) an X-Ray powder diffraction (XRPD) pattern substantially the    same as shown in FIG. 10;-   (b) an X-ray powder diffraction (XRPD) pattern with characteristic    peaks at 8.6±0.1° 2-Theta, 19.3±0.1° 2-Theta, 20.8±0.1° 2-Theta,    24.3±0.1° 2-Theta, 27.6±0.1° 2-Theta;-   (c) a DSC thermogram substantially similar to the one set forth in    FIG. 18;-   (d) a thermo-gravimetric analysis (TGA) thermogram substantially    similar to the one set forth in FIG. 18;-   (e) a DSC thermogram with a first endotherm having an onset    temperature at about 104° C. and second endotherm having an onset    temperature at about 193° C.;    or-   (f) combinations thereof.

In some embodiments, Form J is characterized as having at least two, atleast three, at least four, or all least five of the properties selectedfrom (a) to (e).

In some embodiments, Form J is characterized as having an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 10.In some embodiments, Form J is characterized as having an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 8.6±0.1°2-Theta, 19.3±0.1° 2-Theta, 20.8±0.1° 2-Theta, 24.3±0.1° 2-Theta,27.6±0.1° 2-Theta.

In some embodiments, Form J is characterized as having a DSC thermogramsubstantially similar to the one set forth in FIG. 18. In someembodiments, Form J is characterized as having a thermo-gravimetricanalysis (TGA) thermogram substantially similar to the one set forth inFIG. 18. In some embodiments, Form J is characterized as having a DSCthermogram with an endotherm having an onset temperature at about 104°C. In some embodiments, Form J is characterized as having a DSCthermogram with a first endotherm having an onset temperature at about104° C. and second endotherm having an onset temperature at about 193°C.

In some embodiments, Form J is obtained from a mixture of acetone andwater. In some embodiments, Form J is solvated. In some embodiments,Form J is an acetone solvate.

Preparation of Crytalline Forms

In some embodiments, crystalline forms of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide are prepared as outlined in the Examples. It is notedthat solvents, temperatures and other reaction conditions presentedherein may vary.

Suitable Solvents

Therapeutic agents that are administrable to mammals, such as humans,must be prepared by following regulatory guidelines. Such governmentregulated guidelines are referred to as Good Manufacturing Practice(GMP). GMP guidelines outline acceptable contamination levels of activetherapeutic agents, such as, for example, the amount of residual solventin the final product. Preferred solvents are those that are suitable foruse in GMP facilities and consistent with industrial safety concerns.Categories of solvents are defined in, for example, the InternationalConference on Harmonization of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH), “Impurities: Guidelines forResidual Solvents, Q3C(R3), (November 2005).

Solvents are categorized into three classes. Class 1 solvents are toxicand are to be avoided. Class 2 solvents are solvents to be limited inuse during the manufacture of the therapeutic agent. Class 3 solventsare solvents with low toxic potential and of lower risk to human health.Data for Class 3 solvents indicate that they are less toxic in acute orshort-term studies and negative in genotoxicity studies.

Class 1 solvents, which are to be avoided, include: benzene; carbontetrachloride; 1,2-dichloroethane; 1,1-dichloroethene; and1,1,1-trichloroethane.

Examples of Class 2 solvents are: acetonitrile, chlorobenzene,chloroform, cyclohexane, 1,2-dichloroethene, dichloromethane,1,2-dimethoxyethane, N,N-dimethylacetamide, N,N-dimethylformamide,1,4-dioxane, 2-ethoxyethanol, ethyleneglycol, formamide, hexane,methanol, 2-methoxyethanol, methylbutyl ketone, methylcyclohexane,N-methylpyrrolidine, nitromethane, pyridine, sulfolane, tetralin,toluene, 1,1,2-trichloroethene and xylene.

Class 3 solvents, which possess low toxicity, include: acetic acid,acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert-butylmethylether (MTBE), cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethylether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropylacetate, methyl acetate, 3-methyl-1-butanol, methylethyl ketone,methylisobutyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol,1-propanol, 2-propanol, propyl acetate, and tetrahydrofuran.

In some embodiments, compositions comprising4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideinclude a residual amount of an organic solvent(s). In some embodiments,compositions comprising4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideinclude a detectable amount of an organic solvent(s). In someembodiments, compositions comprising4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideinclude a residual amount of a Class 3 solvent. In some embodiments, theorganic solvent is a Class 3 solvent. In some embodiments, the Class 3solvent is selected from the group consisting of acetic acid, acetone,anisole, 1-butanol, 2-butanol, butyl acetate, tert-butylmethyl ether,cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethylformate, formic acid, heptane, isobutyl acetate, isopropyl acetate,methyl acetate, 3-methyl-1-butanol, methylethyl ketone, methylisobutylketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol,2-propanol, propyl acetate, and tetrahydrofuran. In some embodiments,the Class 3 solvent is ethanol.

The methods and compositions described herein include the use ofcrystalline forms of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In addition, the crystalline forms of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidedescribed herein can exist in unsolvated as well as solvated forms withpharmaceutically acceptable solvents such as water, ethanol, and thelike.

Definitions

The term “pharmaceutically acceptable excipient,” as used herein, refersto a material, such as a carrier, diluent, stabilizer, dispersing agent,suspending agent, thickening agent, etc. which allows processing theactive pharmaceutical ingredient (API) into a form suitable foradministration to a mammal. In one aspect, the mammal is a human.Pharmaceutically acceptable excipients refer to materials which do notsubstantially abrogate the desired biological activity or desiredproperties of the compound (i.e. API), and is relatively nontoxic, i.e.,the material is administered to an individual without causingundesirable biological effects or interacting in a deleterious mannerwith any of the components of the composition in which it is contained.

“Active pharmaceutical ingredient” or API refers to a compound thatpossesses a desired biological activity or desired properties. In someembodiments, an API is4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In some embodiments, an API is crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.In some embodiments, the API has a purity of greater than 90%, greaterthan 95%, greater than 96%, greater than 97%, greater than 98%, greaterthan 98%, or greater than 99%.

The term “pharmaceutical composition” refers to a mixture of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or pharmaceutically acceptable salt and/or solvate thereof, with otherchemical components, such as carriers, stabilizers, diluents, dispersingagents, suspending agents, thickening agents, excipients, etc. Thepharmaceutical composition facilitates administration of the compound toa mammal.

Administration of a combination of agents, as used herein, includesadministration of the agents described in a single composition or in acombination therapy wherein one or more agent is administered separatelyfrom at least one other agent.

“Detectable amount” refers to an amount that is measurable usingstandard analytic methods (e.g. ion chromatography, mass spectrometry,NMR, HPLC, gas chromatography, elemental analysis, IR spectroscopy,inductively coupled plasma atomic emission spectrometry, USP<231>MethodII, etc) (ICH guidances, Q2A Text on Validation of Analytical Procedures(March 1995) and Q2 B Validation of Analytical Procedures: Methodology(November 1996)).

The term “acceptable” with respect to a formulation, composition oringredient, as used herein, means having no persistent detrimentaleffect on the general health of the subject being treated.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent being administeredwhich will relieve to some extent one or more of the symptoms of thedisease or condition being treated. The result can be reduction and/oralleviation of the signs, symptoms, or causes of a disease, or any otherdesired alteration of a biological system. For example, an “effectiveamount” for therapeutic uses is the amount of the composition comprisinga compound as disclosed herein required to provide a clinicallysignificant decrease in disease symptoms. The term “therapeuticallyeffective amount” includes, for example, a prophylactically effectiveamount. The effective amount will be selected based on the particularpatient and the disease level. It is understood that “an effect amount”or “a therapeutically effective amount” varies from subject to subject,due to variation in metabolism of drug, age, weight, general conditionof the subject, the condition being treated, the severity of thecondition being treated, and the judgment of the prescribing physician.In one embodiment, an appropriate “effective” amount in any individualcase is determined using techniques, such as a dose escalation study

The terms “enhance” or “enhancing,” as used herein, means to increase orprolong either in potency or duration a desired effect. Thus, in regardto enhancing the effect of therapeutic agents, the term “enhancing”refers to the ability to increase or prolong, either in potency orduration, the effect of other therapeutic agents on a system. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of another therapeutic agent in a desiredsystem.

The terms “kit” and “article of manufacture” are used as synonyms.

The term “modulate,” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

The term “modulator” as used herein, refers to a molecule that interactswith a target either directly or indirectly. The interactions include,but are not limited to, the interactions of an agonist, partial agonist,an inverse agonist, antagonist, degrader, AR trafficking modulator, ARDNA-binding inhibitor. In some embodiments, a modulator is anantagonist. In some embodiments, a modulator is an inverse agonist,antagonist, degrader, AR trafficking modulator and/or a DNA bindinginhibitor.

The term “antagonist” as used herein, refers to a small-molecule agentthat binds to a nuclear hormone receptor and subsequently decreases theagonist induced transcriptional activity of the nuclear hormonereceptor.

The term “agonist” as used herein, refers to a small-molecule agent thatbinds to a nuclear hormone receptor and subsequently increases nuclearhormone receptor transcriptional activity in the absence of a knownagonist.

The term “inverse agonist” as used herein, refers to a small-moleculeagent that binds to a nuclear hormone receptor and subsequentlydecreases the basal level of nuclear hormone receptor transcriptionalactivity that is present in the absence of a known agonist.

The term “degrader” as used herein, refers to a small molecule agentthat binds to a nuclear hormone receptor and subsequently lowers thesteady state protein levels of said receptor.

The term “AR trafficking modulator” as used herein, refers to asmall-molecule agent that binds to a nuclear hormone receptor andsubsequently alters the normal subcellular location of the receptorthereby interfering with its function and signaling.

The term “DNA-binding inhibitor” as used herein, refers to asmall-molecule agent that binds to a nuclear hormone receptor andsubsequently prevents DNA binding of the receptor thereby interferingwith its function and signaling.

“Selective” with respect to androgen receptors means that the compoundpreferentially binds to androgen receptors versus other nuclearreceptors. In some embodiments, a selective androgen receptor modulatorpreferentially binds to androgen receptors and displays little, if any,affinity to other nuclear receptors.

The term “cancer” as used herein refers to an abnormal growth of cellswhich tend to proliferate in an uncontrolled way and, in some cases, tometastasize (spread).

The term “subject” or “patient” encompasses mammals. In one aspect, themammal is a human. In another aspect, the mammal is a non-human primatesuch as chimpanzee, and other apes and monkey species. In one aspect,the mammal is a farm animal such as cattle, horse, sheep, goat, orswine. In one aspect, the mammal is a domestic animal such as rabbit,dog, or cat. In one aspect, the mammal is a laboratory animal, includingrodents, such as rats, mice and guinea pigs, and the like.

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating at least one symptom of a diseaseor condition, preventing additional symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Pharmaceutical Compositions/Formulations

Pharmaceutical compositions are formulated in a conventional mannerusing one or more physiologically acceptable carriers comprisingexcipients and auxiliaries which facilitate processing of the activecompounds into preparations which are used pharmaceutically. Suitabletechniques, carriers, and excipients include those found within, forexample, Remington: The Science and Practice of Pharmacy, Nineteenth Ed(Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E.,Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa.1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins 1999), herein incorporated by reference in their entirety.

In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis formulated for oral administration to a mammal. In some embodiments,crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis formulated into an oral dosage form. In some embodiments, crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis formulated into a solid oral dosage form. In some embodiments,crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis formulated into a tablet, powder, pill, capsule, and the like, fororal ingestion by a mammal.

Contemplated pharmaceutical compositions provide a therapeuticallyeffective amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideenabling, for example, once-a-day, twice-a-day, three times a day, etc.administration. In one aspect, pharmaceutical compositions provide aneffective amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideenabling once-a-day dosing.

Dose Amounts

In certain embodiments, the amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein the pharmaceutical compositions is about 0.3 mg to about 1.5 g perdose, 0.3 mg to about 1 g per dose, about 1 mg to about 1 g per dose.

In one embodiment, the amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein the pharmaceutical compositions is about 1 mg per dose, about 5 mgper dose, about 10 mg per dose, about 15 mg per dose, about 30 mg perdose, about 45 mg per dose, about 60 mg per dose, about 100 mg per dose,about 150 mg per dose, about 200 mg per dose, about 300 mg per dose,about 400 mg per dose, about 500 mg per dose, about 600 mg per dose, orabout 1000 mg per dose. In some embodiments, the amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein the pharmaceutical compositions is about 30 mg per dose. In someother embodiments, the amount of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein the pharmaceutical compositions is about 60 mg per dose.

In general, doses employed for adult human treatment are typically inthe range of 0.01 mg-5000 mg per day. In one aspect, doses employed foradult human treatment are from about 1 mg to about 1000 mg per day. Insome embodiments, doses employed for adult human treatment are about 240mg per day. In one embodiment, the desired dose is convenientlypresented in a single dose or in divided doses administeredsimultaneously (or over a short period of time) or at appropriateintervals, for example as two, three, four or more sub-doses per day.

In one embodiment, the daily dosages appropriate for4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis from about 0.01 to about 20 mg/kg per body weight. In otherembodiments, the daily dosage or the amount of active in the dosage formare lower or higher than the ranges indicated herein.

Methods of Dosing and Treatment Regimens

In one embodiment, the pharmaceutical compositions including4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidedescribed herein are administered for prophylactic and/or therapeutictreatments. In therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest at leastone of the symptoms of the disease or condition. In certain embodiments,amounts effective for this use depend on the severity and course of thedisease or condition, previous therapy, the patient's health status,weight, and response to the drugs, and/or the judgment of the treatingphysician.

In prophylactic applications, compositions containing4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidedescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition. Suchan amount is defined to be a “prophylactically effective amount ordose.” In this use, the precise amounts also depend on the patient'sstate of health, weight, and the like. When used in a patient, effectiveamounts for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician.

In certain embodiments, administration of the compound, compositions ortherapies as described herein includes chronic administration. Incertain embodiments, chronic administration includes administration foran extended period of time, including, e.g., throughout the duration ofthe patient's life in order to ameliorate or otherwise control or limitthe symptoms of the patient's disease or condition. In some embodiments,chronic administration includes daily administration.

In some embodiments, administration of the compound, compositions ortherapies described herein is given continuously. In alternativeembodiments, the dose of drug being administered is temporarily reducedor temporarily suspended for a certain length of time (i.e., a “drugholiday”). The length of the drug holiday varies between 2 days and 1year, including by way of example only, 2 days, 3 days, 4 days, 5 days,6 days, 7 days, 10 days, 12 days, 15 days, 20 days, 28 days, 35 days, 50days, 70 days, 100 days, 120 days, 150 days, 180 days, 200 days, 250days, 280 days, 300 days, 320 days, 350 days, and 365 days. The dosereduction during a drug holiday is from 10%-100%, including by way ofexample only 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, and 100%.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, in specificembodiments, the dosage or the frequency of administration, or both, isreduced, as a function of the symptoms, to a level at which the improveddisease, disorder or condition is retained. In certain embodiments,however, the patient requires intermittent treatment on a long-termbasis upon any recurrence of symptoms.

The amount of a given agent that corresponds to such an amount variesdepending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight, sex) of thesubject or host in need of treatment, but can nevertheless be determinedaccording to the particular circumstances surrounding the case,including, e.g., the specific agent being administered, the route ofadministration, the condition being treated, and the subject or hostbeing treated.

Combination Treatments

In certain instances, it is appropriate to administer4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein combination with another therapeutic agent.

In one embodiment, the compositions and methods described herein arealso used in conjunction with other therapeutic reagents that areselected for their particular usefulness against the condition that isbeing treated. In general, the compositions described herein and, inembodiments where combinational therapy is employed, other agents do nothave to be administered in the same pharmaceutical composition, and are,because of different physical and chemical characteristics, administeredby different routes. In one embodiment, the initial administration ismade according to established protocols, and then, based upon theobserved effects, the dosage, modes of administration and times ofadministration, further modified.

In various embodiments, the compounds are administered concurrently(e.g., simultaneously, essentially simultaneously or within the sametreatment protocol) or sequentially, depending upon the nature of thedisease, the condition of the patient, and the actual choice ofcompounds used. In certain embodiments, the determination of the orderof administration, and the number of repetitions of administration ofeach therapeutic agent during a treatment protocol, is based uponevaluation of the disease being treated and the condition of thepatient.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drugemployed, on the specific drug employed, on the disease or conditionbeing treated and so forth.

The individual compounds of such combinations are administered eithersequentially or simultaneously in separate or combined pharmaceuticalformulations. In one embodiment, the individual compounds will beadministered simultaneously in a combined pharmaceutical formulation.Appropriate doses of known therapeutic agents will be appreciated bythose skilled in the art.

The combinations referred to herein are conveniently presented for usein the form of a pharmaceutical compositions together with apharmaceutically acceptable diluent(s) or carrier(s).

Kits/Articles of Manufacture

For use in the therapeutic methods of use described herein,kits/articles of manufacture are also described herein. Such kitsinclude a carrier, package, or container that is optionallycompartmentalized to receive one or more doses of a pharmaceuticalcomposition of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidefor use in a method described herein. The kits provided herein containpackaging materials. Packaging materials for use in packagingpharmaceutical products include, but are not limited to those describedin e.g., U.S. Pat. No. 5,323,907. Examples of pharmaceutical packagingmaterials include, but are not limited to, blister packs, bottles,tubes, bags, containers, bottles, and any packaging material suitablefor a selected formulation and intended mode of administration andtreatment. A wide array of formulations of the compounds andcompositions provided herein are contemplated as are a variety oftreatments for any disease, disorder, or condition that would benefit bytreatment with an AR antagonist.

For example, the container(s) include4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or a pharmaceutically acceptable salt thereof, optionally in acomposition or in combination with another agent as disclosed herein.Such kits optionally include an identifying description or label orinstructions relating to its use in the methods described herein.

A kit typically includes labels listing contents and/or instructions foruse, and package inserts with instructions for use. A set ofinstructions will also typically be included.

In one embodiment, a label is on or associated with the container. Inone embodiment, a label is on a container when letters, numbers or othercharacters forming the label are attached, molded or etched into thecontainer itself; a label is associated with a container when it ispresent within a receptacle or carrier that also holds the container,e.g., as a package insert. In one embodiment, a label is used toindicate that the contents are to be used for a specific therapeuticapplication. The label also indicates directions for use of thecontents, such as in the methods described herein.

In certain embodiments, the pharmaceutical compositions are presented ina pack or dispenser device which contains one or more unit dosage formscontaining a compound provided herein. The pack, for example, containsmetal or plastic foil, such as a blister pack. In one embodiment, thepack or dispenser device is accompanied by instructions foradministration. In one embodiment, the pack or dispenser is alsoaccompanied with a notice associated with the container in formprescribed by a governmental agency regulating the manufacture, use, orsale of pharmaceuticals, which notice is reflective of approval by theagency of the form of the drug for human or veterinary administration.Such notice, for example, is the labeling approved by the U.S. Food andDrug Administration for prescription drugs, or the approved productinsert. In one embodiment, compositions containing a compound providedherein formulated in a compatible pharmaceutical carrier are alsoprepared, placed in an appropriate container, and labeled for treatmentof an indicated condition.

EXAMPLES

The following ingredients, formulations, processes and procedures forpracticing the methods disclosed herein correspond to that describedabove. The procedures below describe with particularity illustrative,non-limiting embodiment of formulations that include a4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,or a pharmaceutically acceptable salt and/or solvate thereof, andpharmacokinetic profiles and pharmacodynamic effects thereof. By way ofexample only,4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideis optionally prepared as outlined in U.S. patent application Ser. No.12/294,881, U.S. patent application Ser. No. 12/450,423 or as outlinedherein.

Example 1: Preparation of Crystalline Forms of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide

Form A

2 volumes of ethanol were added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(180 mg). After 6 days the material was filtered. The sample was placedin an oven at 35° C. and ca. 40 mbar pressure for an hour. The isolatedmaterial was shown to be an ethanol solvate by TGA, DSC, GVS and ¹H NMRanalysis. Under forcing conditions (60° C. at <20 mm Hg for 8 days),Form A lost ethanol, the XRPD pattern of the material stayed the same.

Alternatively, THF (1 volume), DCM (1 volume), acetone (1 volume),ethanol (1 volume), methanol (1 volume), nitromethane (1 volume), water(1 volume+sonication), THF-water mixture (1 volume), or dioxane-watermixture (1 volume) was added to approximately 65 mg of the amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The samples were left in screw capped vials at ambient conditions for 3days. Lids on the samples which showed no precipitation were loosened toallow for slow evaporation of the solvent. After a day, these sampleswere placed in a maturation chamber, the temperature of which wasswitched between room temperature and 50° C. every 4 hours. Solidmaterial was isolated. The single crystal XRD studies of Form A(obtained from methanol) confirmed that Form A was a disordered,solvated, hydrated crystalline form and therefore represented a group ofisostructural solvates.

Form B

10 volumes of water were added to crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(Form A; 500 mg). The resulting mixture was stirred for 18 hours at 55°C. The solid was cooled to room temperature. The sample was filtered andwashed using 5 volumes of water. The solid was dried in an oven at 40°C. and ca. 55 mbar pressure for 24 hours.

Alternatively, 5 volumes of ethyl acetate was added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(250 mg) and the resulting solution was placed in a maturation chamber(switched between room temperature and 50° C. every 4 hours) for 5 days.No solid was recovered and some additions amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidewas added until some precipitate appeared. The solution was left tostand at room temperature to allow slow evaporation of the solution.After 6 days the solid was filtered and dried in an oven at 35° C. andca. 40 mbar for an hour.

In another embodiment, approximately 10 mg of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(Form A) was transferred to a HPLC vial. A solution was prepared byadding gradually TBME (400 μL) or toluene (800 μL) to the material.After each successive 200 μl addition, the vial was shaken at 50° C. tohelp dissolution. Once a clear solution was obtained, the vial was leftto stand at room temperature with the septum pierced with a needle toallow for slow evaporation of the solvent. After 2 weeks cube-likecrystals were obtained from toluene and were submitted for singlecrystal X-ray diffraction (SCXRD) (see Example 4). The crystallinestructure was solved and the form was found to be an unsolvatedcrystalline form (Form B).

In yet another embodiment, toluene (2 volume), isopropylacetate (2volume) or MEK (1 volume) was added to approximately 65 mg of theamorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The samples were left in screw capped vials at ambient conditions for 3days. Lids on the samples which showed no precipitation were loosened toallow for slow evaporation of the solvent. After a day, these sampleswere placed in a maturation chamber, the temperature of which wasswitched between room temperature and 50° C. every 4 hours. Solidmaterial was isolated.

Form C

4 volumes of isopropanol were added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(160 mg). After 6 days the material was filtered. The sample was placedin an oven at 35° C. and ca. 40 mbar pressure for an hour.

Alternatively, anisole (2 volume), IPA (1 volume) or IPA-water mixture(1 volume) were added to approximately 65 mg of the amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The samples were left in screw capped vials at ambient conditions for 3days. Lids on the samples which showed no precipitation were loosened toallow for slow evaporation of the solvent. After a day, these sampleswere placed in a maturation chamber, the temperature of which wasswitched between room temperature and 50° C. every 4 hours. Solidmaterial was isolated.

Form D

5 volumes of MTBE was added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(200 mg) and the resulting mixture was placed in a maturation chamber(switched between room temperature and 50° C. every 4 hours) for 5 days.The solid obtained was filtered and dried in an oven at 35° C. and ca.40 mbar pressure for an hour.

Alternatively, MTBE (2 volumes) was added to approximately 65 mg of theamorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The sample was left in screw capped vial at ambient conditions for 3days. If the sample showed no precipitation, the lid was loosened toallow for slow evaporation of the solvent. After a day, this sample wasplaced in a maturation chamber, the temperature of which was switchedbetween room temperature and 50° C. every 4 hours. Solid material wasisolated.

Form E

DMSO (1 volume) was added to approximately 65 mg of the amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The sample was left in screw capped vial at ambient conditions for 3days. If the sample showed no precipitation, the lid was loosened toallow for slow evaporation of the solvent. After a day, this sample wasplaced in a maturation chamber, the temperature of which was switchedbetween room temperature and 50° C. every 4 hours. Solid material wasisolated.

Form F

An acetone/water mixture (1 volume) was added to approximately 65 mg ofthe amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The sample was left in screw capped vial at ambient conditions for 3days. If the sample showed no precipitation, the lid was loosened toallow for slow evaporation of the solvent. After a day, this sample wasplaced in a maturation chamber, the temperature of which was switchedbetween room temperature and 50° C. every 4 hours. Solid material wasisolated.

Under ambient conditions, within a month, Form F transformed to Form A.

Form G

4 volumes of 2-methoxyethanol were added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(160 mg). After 6 days the material was filtered. The solid was placedin an oven at 35° C. and ca. 40 mbar pressure for an hour.

Alternatively, 2-methoxyethanol (1 volume) was added to approximately 65mg of the amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The sample was left in screw capped vial at ambient conditions for 3days. If the sample showed no precipitation, the lid was loosened toallow for slow evaporation of the solvent. After a day, this sample wasplaced in a maturation chamber, the temperature of which was switchedbetween room temperature and 50° C. every 4 hours. Solid material wasisolated.

Form H

Ethyl acetate (2 volumes) was added to approximately 65 mg of theamorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.A minimum amount of solvent was added just to wet the material (visuallythis meant softening of the amorphous solid, referred to as collapse).The sample was left in screw capped vial at ambient conditions for 3days. If the sample showed no precipitation, the lid was loosened toallow for slow evaporation of the solvent. After a day, these sampleswere placed in a maturation chamber, the temperature of which wasswitched between room temperature and 50° C. every 4 hours. Solidmaterial was isolated.

Form I

2 volumes of DMSO were added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(150 mg). After 6 days, two lumps of material were obtained one yellowand the other white colored. The yellow colored material was Form E andthe white colored material exhibited a new XRPD. The white coloredmaterial was designated as Form I.

Form J

1.9 volumes of acetone and 0.1 volumes of water were added to amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide(200 mg). The lid was left loose and after 6 days the material was foundto be completed dry. The resulting material was designated as Form J.

Example 2: Preparation of Amorphous4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide

10 Volumes of dichloromethane was added to crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidefollowed by sonication at 48° C. to provide a clear solution. Theresulting solution was rotary evaporated for an hour leading to completeamorphisation of the material (as verified by XRPD analysis).

Example 3: X-Ray Powder Diffraction (XRPD)

X-Ray powder diffraction patterns were collected on a Bruker AXS C2GADDS or a Bruker AXS D8 Advance diffractometer.

Bruker AXS C2 GADDS

X-Ray Powder Diffraction patterns were collected on a Bruker AXS C2GADDS diffractometer using Cu Ka radiation (40 kV, 40 mA), automated XYZstage, laser video microscope for auto-sample positioning and a HiStar2-dimensional area detector. X-ray optics consists of a single Göbelmultilayer mirror coupled with a pinhole collimator of 0.3 mm. The beamdivergence, i.e. the effective size of the X-ray beam on the sample, wasapproximately 4 mm. A θ-θ continuous scan mode was employed with asample detector distance of 20 cm which gives an effective 2θ range of3.2°-29.7°. Typically the sample would be exposed to the X-ray beam for120 seconds. The software used for data collection was GADDS for WNT4.1.16 and the data were analyzed and presented using Diffrac Plus EVA v9.0.0.2 or v 13.0.0.2.

Samples run under ambient conditions were prepared as flat platespecimens using powder as received without grinding. Approximately 1-2mg of the sample was lightly pressed on a glass slide to obtain a flatsurface.

Samples run under non-ambient conditions were mounted on a silicon waferwith heat-conducting compound. The sample was then heated to theappropriate temperature at ca. 10° C.min⁻¹ and subsequently heldisothermally for ca 1 minute before data collection was initiated.

Bruker AXS D8 Advance

X-Ray Powder Diffraction patterns were collected on a Bruker D8diffractometer using Cu Ka radiation (40 kV, 40 mA), θ-2θ goniometer,and divergence of V4 and receiving slits, a Ge monochromator and aLynxeye detector. The instrument is performance checked using acertified Corundum standard (NIST 1976). The software used for datacollection was Diffrac Plus XRD Commander v2.5.0 and the data wereanalyzed and presented using Diffrac Plus EVA v 11,0.0.2 or v 13.0.0.2.Samples were run under ambient conditions as flat plate specimens usingpowder. Approximately 20 mg of the sample was gently packed into acavity cut into polished, zero-background (510) silicon wafer. Thesample was rotated in its own plane during analysis. The details of thedata collection are:

-   -   Angular range: 2 to 42° 2θ    -   Step size: 0.05° 2θ    -   Collection time: 0.5 s.step⁻¹

Form A

The X-Ray powder diffraction pattern for Form A is displayed in FIG. 1.Characteristic peaks include 4.8±0.1° 2-Theta, 7.1±0.1° 2-Theta,14.2±0.1° 2-Theta, 16.3±0.1° 2-Theta, 20.1±0.1° 2-Theta.

Form B

The X-Ray powder diffraction pattern for Form B is displayed in FIG. 2.Characteristic peaks include 12.1±0.1° 2-Theta, 16.0±0.1° 2-Theta,16.7±0.1° 2-Theta, 20.1±0.1° 2-Theta, 20.3±0.1° 2-Theta.

Form C

The X-Ray powder diffraction pattern for Form C is displayed in FIG. 3.Characteristic peaks include 4.3±0.1° 2-Theta, 6.9±0.1° 2-Theta,9.1±0.1° 2-Theta, 10.6±0.1° 2-Theta, 13 8±0.1° 2-Theta, 26.4±0.1°2-Theta.

Form D

The X-Ray powder diffraction pattern for Form D is displayed in FIG. 4.Characteristic peaks include 6.3±0.1° 2-Theta, 13.9±0.1° 2-Theta,16.4±0.1° 2-Theta, 17.0±0.1° 2-Theta, 23.7±0.1° 2-Theta, 24.8±0.1°2-Theta.

Form E

The X-Ray powder diffraction pattern for Form E is displayed in FIG. 5.Characteristic peaks include 7.2±0.1° 2-Theta, 11.8±0.1° 2-Theta,16.1±0.1° 2-Theta, 20.5±0.1° 2-Theta, 23.0±0.1° 2-Theta, 25.2±0.1°2-Theta. Variable temperature XRPD showed transformation of Form E toForm A to Form B.

Form F

The X-Ray powder diffraction pattern for Form F is displayed in FIG. 6.Characteristic peaks include 4.6±0.1° 2-Theta, 6.1±0.1° 2-Theta,14.3±0.1° 2-Theta, 21.6±0.1° 2-Theta, 22.4±0.1° 2-Theta, 23.3±0.1°2-Theta, 25.5±0.1° 2-Theta.

Form G

The X-Ray powder diffraction pattern for Form G is displayed in FIG. 7.Characteristic peaks include 7.0±0.1° 2-Theta, 10.3±0.1° 2-Theta,14.1±0.1° 2-Theta, 15.2±0.1° 2-Theta, 23.6±0.1° 2-Theta.

Form H

The X-Ray powder diffraction pattern for Form H is displayed in FIG. 8.Characteristic peaks include 8.0±0.1° 2-Theta, 14.7±0.1° 2-Theta,15.9±0.1° 2-Theta, 18.2±0.1° 2-Theta, 25.7±0.1° 2-Theta, 26.7±0.1°2-Theta.

Form I

The X-Ray powder diffraction pattern for Form I is displayed in FIG. 9.Characteristic peaks include 7.7±0.1° 2-Theta, 10.4±0.1° 2-Theta,11.6±0.1° 2-Theta, 17.0±0.1° 2-Theta, 20.0±0.1° 2-Theta, 20.6±0.1°2-Theta.

Form J

The X-Ray powder diffraction pattern for Form J is displayed in FIG. 10.Characteristic peaks include 8.6±0.1° 2-Theta, 19.3±0.1° 2-Theta,20.8±0.1° 2-Theta, 24.3±0.1° 2-Theta, 27.6±0.1° 2-Theta.

Example 4: Single Crystal X-Ray Diffraction (SCXRD)

Single crystal X-ray diffraction data was collected on an OxfordDiffraction Supernova Dual Source, Cu at Zero, Atlas CCD diffractometerequipped with an Oxford Cryosystems Cryostream/Cobra cooling device. Thedata was collected using CuKα/MoKα radiation. Structures were typicallysolved using either the SHELXS or SHELXD programs and refined with theSHELXL program as part of the Bruker AXS SHELXTL suite. Unless otherwisestated, hydrogen atoms attached to carbon were placed geometrically andallowed to refine with a riding isotropic displacement parameter.Hydrogen atoms attached to a heteroatom were located in a differenceFourier synthesis and were allowed to refine freely with an isotropicdisplacement parameter.

Form A

Form A is characterized by unit cell unit cell parameters approximatelyequal to the following at a temperature of approximately −173° C.:

TABLE 1 Single Crystal Structure of Form A Molecular formulaC₂₁H₁₅F₄N₅O₂S₁ Molecular weight 485.5 Crystal system Orthorhombic Spacegroup P2(1)2(1)2 a 16.3429(3) Å α 90° b 37.7298(7) Å β 90° c 7.23410(10)Å  γ 90° V 4460.65(13) Å3 Z 8 Dc 1.446 g · cm⁻¹ μ 0.207 mm⁻¹ Source, λMo—K(alpha), 0.71073 Å F(000) 2016 T 100(2)K Crystal Colourless block,0.25 × 0.2 × 0.1 mm Data truncated to 0.80 Å θmax 26.37° Completeness99.4% Reflections 67442 Unique reflections 9056 Rint 0.0425

The structure solution was obtained by direct methods, full-matrixleast-squares refinement on F² with weighting w⁻¹=σ2(F_(o)²)+(0.1070P)²+(6.5000P), where P=(F_(o) ²+2F_(c) ²)/3, anisotropicdisplacement parameters, empirical absorption correction using sphericalharmonics, implemented in SCALE3 ABSPACK scaling algorithm. FinalwR²={Σ[w(F_(o) ²−F_(c) ²)]/Σ[w(F_(o) ²)²]^(1/2)}=0.1814 for all data,conventional R₁=0.0652 on F values of 7570 reflections withF_(o)>4σ(F_(o)), S=1.005 for all data and 642 parameters. Final Δ/σ(max)0.004, Δ/σ (mean), 0.000. Final difference map between +1.158 and−0.443 e Å⁻³.

A simulated XRPD obtained from the single crystal data for Form Amatched the experimental XRPD.

The single crystal XRD analysis confirmed that Form A is a disordered,solvated, hydrated crystalline form. Since Form A was obtained fromdifferent solvents, it can be concluded that Form A represents a groupof isostructural solvates.

Form B

Form B is characterized by unit cell unit cell parameters approximatelyequal to the following at a temperature of approximately −173° C.:

TABLE 2 Single Crystal Structure of Form B Molecular formulaC₂₁H₁₅F₄N₅O₂S Molecular weight 477.44 Crystal system Monoclinic Spacegroup P2₁/c a 17.7796(4) Å α  90° b 12.9832(3) Å β 100.897(2)° c18.4740(4) Å γ  90° V 4187.57(16) Å³ Z 8 Dc 1.515 g · cm⁻¹ μ 0.22 mm⁻¹Source, λ Mo—K(alpha), 0.71073 Å F(000) 1952 T 100(2)K Crystalcolourless prism, 0.23 × 0.20 × 0.05 mm,, 0.3 × 0.3 × 0.2 mm Datatruncated to 0.80 Å θmax 26.37° Completeness 99.6% Reflections 27616Unique reflections 8527 Rint 0.0458

The structure solution was obtained by direct methods, full-matrixleast-squares refinement on F² with weighting w⁻¹=σ2(F_(o)²)+(0.0425P)²+(0.0000P), where P=(F_(o) ²+2F_(c) ²)/3, anisotropicdisplacement parameters, empirical absorption correction using sphericalharmonics, implemented in SCALE3 ABSPACK scaling algorithm. FinalwR²={Σ[w(F_(o) ²−F_(c) ²)²]/Σ[w(F_(o) ²)²]^(1/2)}=0.0941 for all data,conventional R₁=0.0404 on F values of 5767 reflections withF_(o)>4σ(F_(o)), S=1.005 for all data and 613 parameters. Final Δ/σ(max) 0.001, Δ/σ (mean), 0.000. Final difference map between +0.76 and−0.603 e Å⁻³.

A simulated XRPD obtained from the single crystal data for Form Bmatched the experimental XRPD.

The single crystal XRD analysis confirmed that Form B is unsolvated.

Form E

Form E is characterized by unit cell unit cell parameters approximatelyequal to the following at a temperature of approximately −173° C.:

TABLE 3 Single Crystal Structure of Form E Molecular formulaC₂₃H₂₁F₄N₅O₃S₂ Molecular weight 555.57 Crystal system Orthorhombic Spacegroup P_(na)2₁ a 8.43080(10) Å  α 90° b 17.1685(3) Å β 90° c 17.4276(3)Å γ 90° V 2522.54(7) Å³ Z 4 Dc 1.463 g · cm⁻¹ μ 2.504 mm⁻¹ Source, λ CuKα, 1.54178 Å F(000) 1144 T 100(2)K Crystal colourless prism, 0.23 ×0.20 × 0.05 mm,, 0.3 × 0.2 × 0.07 mm Data truncated to 0.80 Å θmax74.48° Completeness 99.6% Reflections 11318 Unique reflections 4424 Rint0.019

The structure solution was obtained by direct methods, full-matrixleast-squares refinement on F² with weighting w⁻¹=σ2(F_(o)²)+(0.1120P)²+(1.1000P), where P=(F_(o) ²+2F_(c) ²)/3, anisotropicdisplacement parameters, empirical absorption correction using sphericalharmonics, implemented in SCALE3 ABSPACK scaling algorithm. FinalwR²={Σ[w(F_(o) ²−F_(c) ²)²]/Σ[w(F_(o) ²)²]^(1/2)}=0.1442 for all data,conventional R₁=0.0492 on F values of 4257 reflections withF_(o)>4σ(F_(o)), S=1.01 for all data and 342 parameters. Final Δ/σ (max)0.000, Δ/σ (mean), 0.000. Final difference map between +1.923 and −0.527e Å⁻³.

A simulated XRPD obtained from the single crystal data for Form Ematched the experimental XRPD.

The single crystal XRD (SCXRD) studies of Form E confirmed that it was a1:1 DMSO solvate.

Form G

Form G is characterized by unit cell unit cell parameters approximatelyequal to the following at a temperature of approximately −173° C.:

TABLE 4 Single Crystal Structure of Form G Molecular formulaC₂₄H₂₃F₄N₅O₄S Molecular weight 553.53 Crystal system Monoclinic Spacegroup Cc a 18.613(2) Å  α 90° b 16.9728(14) Å β 91.328(8)° c 7.8214(7) Åγ 90° V 2470.2(4) Å³ Z 4 Dc 1.488 g · cm⁻¹ μ 0.203 mm⁻¹ Source, λMo—K(alpha), 0.71073 Å F(000) 1144 T 100(2)K Crystal colourless prism,0.23 × 0.20 × 0.05 mm,, 0.5 × 0.1 × 0.1 mm Data truncated to 0.80 Å θmax26.37° Completeness 99.6% Reflections 11648 Unique reflections 4309 Rint0.0565

The structure solution was obtained by direct methods, full-matrixleast-squares refinement on F² with weighting w⁻¹=σ2(F_(o)²)+(0.0790P)²+(0.0000P), where P=(F_(o) ²+2F_(c) ²)/3, anisotropicdisplacement parameters, empirical absorption correction using sphericalharmonics, implemented in SCALE3 ABSPACK scaling algorithm. FinalwR²={Σ[w(F_(o) ²−F_(c) ²)²]/Σ[w(F_(o) ²)²]^(1/2)}=0.114 for all data,conventional R₁=0.0442 on F values of 3799 reflections withF_(o)>4σ(F_(o)), S=1.005 for all data and 353 parameters. Final Δ/σ(max) 0.000, Δ/σ (mean), 0.000. Final difference map between +0.502 and−0.401 e Å⁻³.

A simulated XRPD obtained from the single crystal data for Form Gmatched the experimental XRPD.

The single crystal XRD studies (SCXRD) of Form G confirmed that it was a1:1 2-methoxyethanol solvate.

Example 5: Differential Scanning Calorimetry (DSC) and ThermogravimetricAnalysis (TGA)

DSC data were collected on a TA Instruments Q2000 or Mettler DSC 823e.

In some cases, DSC data were collected on a TA Instruments Q2000equipped with a 50 position autosampler. The calibration for thermalcapacity was carried out using sapphire and the calibration for energyand temperature was carried out using certified indium. Typically 0.5-3mg of each sample, in a pin-holed aluminium pan, was heated at 10°C.min⁻¹ from 25° C. to 350° C. A purge of dry nitrogen at 50 ml.min⁻¹was maintained over the sample. Modulated temperature DSC was carriedout using an underlying heating rate of 2° C.min−1 and temperaturemodulation parameters of ±0.2° C.min⁻¹ and 40 seconds. The instrumentcontrol software was Advantage for Q Series v2.8.0.392 and ThermalAdvantage v4.8.3 and the data were analysed using Universal Analysisv4.3A.

In other cases, DSC data were collected on a Mettler DSC 823e equippedwith a 34 position auto-sampler. The instrument was calibrated forenergy and temperature using certified indium. Typically 0.5-3 mg ofeach sample, in a pin-holed aluminium pan, was heated at 10° C.min⁻¹from 25° C. to 350° C. A nitrogen purge at 50 ml·min⁻¹ was maintainedover the sample. The instrument control and data analysis software wasSTARe v9.20.

TGA data were collected on a TA Instruments Q500 or Mettler TGA/SDTA851e.

In some cases, TGA data were collected on a TA Instruments Q500 TGA,equipped with a 16 position autosampler. The instrument was temperaturecalibrated using certified Alumel. Typically 5-30 mg of each sample wasloaded onto a pre-tared platinum crucible and aluminium DSC pan, and washeated at 10° C.·min⁻¹ from ambient temperature to 350° C. A nitrogenpurge at 60 ml.min−1 was maintained over the sample. The instrumentcontrol software was Advantage for Q Series v2.8.0.392 and ThermalAdvantage v4.8.3.

In other cases, TGA data were collected on a Mettler TGA/SDTA 851eequipped with a 34 position autosampler. The instrument was temperaturecalibrated using certified indium. Typically 5-30 mg of each sample wasloaded onto a pre-weighed aluminium crucible and was heated at 10°C.·min⁻¹ from ambient temperature to 350° C. A nitrogen purge at 50ml·min⁻¹ was maintained over the sample. The instrument control and dataanalysis software was STARe v9.20.

Form A

The single crystal XRD analysis confirmed that Form A is a disordered,solvated, hydrated crystalline form. A sample of the ethanol solvateshowed an endotherm having an onset at about 108-120° C. and a peak atabout 133-135° C. A representative DSC thermogram is shown in FIG. 19.In some embodiments, variable temperature XRPD experiments showed Form Ato become amorphous above about 120° C. followed by a recrystallizationto Form B at about 175° C., which subsequently melted at about 194° C.

Form B

A sample of Form B was analyzed by TGA and DSC and the thermograms areshown in FIG. 11. TGA showed no weight loss above the decompositiontemperature and DSC showed a sharp melting endotherm with an onsettemperature at about 194° C.

Form C

A sample of Form C (from isopropanol) was analyzed by TGA and DSC andthe thermograms are shown in FIG. 12. An endotherm with an onsettemperature at about 118° C. was observed. A small endotherm with anonset temperature at about 193° C. was also observed. The weight lossobserved in the TGA experiment matched the temperature range in whichthe Form lost crystallinity by VT-XRPD suggesting that Form C was notunsolvated. 0.45 equivalents of isopropanol was observed by ¹H NMR and0.49 equivalents of isopropanol was calculated from the weight loss inTGA. Form C obtained from isopropanol is an isopropanol solvate.

Form D

A sample of Form D was analyzed by TGA and DSC, and the thermograms areshown in FIG. 13. An endotherm with an onset temperature at about 122°C. was observed. A smaller second endotherm with an onset temperature atabout 192° C. was also observed.

The weight loss observed in the TGA experiment matched the temperaturerange in which the Form lost crystallinity by VT-XRPD suggesting thatForm D was not unsolvated. 0.26 equivalents of MTBE was observed by ¹HNMR, and 0.26 equivalents of MTBE was calculated from the weight loss inTGA. Form D obtained from MTBE is a MTBE solvate.

Form E

A sample of Form E was analyzed by TGA and DSC, and the thermograms areshown in FIG. 14. A main endotherm having an onset temperature at about116° C. was observed. A relatively small endotherm having an onsettemperature at about 140° C. was also observed. On heating at 10° C./minin a DSC pan, an endotherm at 140° C. was observed. VT-XRPD showedtransformation of Form E to Form A to Form B.

Form F

A sample of Form F was analyzed by TGA and DSC, and the thermograms areshown in FIG. 15. A main endotherm having an onset temperature at about113° C. was observed. A relatively small endotherm having an onsettemperature at about 193° C. was also observed.

Form G

A sample of Form G was analyzed by DSC, and the thermogram is shown inFIG. 16. A main endotherm having an onset temperature at about at 101°C. was observed. A relatively small endotherm having an onsettemperature at about 190° C. was also observed.

Form H

A sample of Form H was analyzed by TGA and DSC, and the thermograms areshown in FIG. 17. The TGA thermogram showed no weight loss below thedecomposition temperature. The DSC thermogram showed a sharp meltingendotherm with an onset temperature of 173° C., and a relatively smallerendotherm with an onset temperature of 193° C. Based on theseobservations and the NMR spectrum (i.e. no significant amount of solventobserved), Form H is unsolvated.

Form J

A sample of Form J was analyzed by TGA and DSC and the thermograms areshown in FIG. 18. An endotherm having an onset temperature at about 104°C. was observed. An endotherm having an onset temperature at about 193°C. was also observed. The weight loss observed in the TGA experimentmatched the temperature range in which Form J lost crystallinity byVT-XRPD suggesting that Form J was not unsolvated. 0.45 equivalents ofacetone was observed by ¹H NMR, and 0.46 equivalents of acetone wascalculated from the weight loss in TGA. Form J obtained from anacetone/water mixture is an acetone solvate.

Example 6: Gravimetric Vapour Sorption (GVS)

Sorption isotherms were obtained using a SMS DVS Intrinsic moisturesorption analyser, controlled by SMS Analysis Suite software. The sampletemperature was maintained at 25° C. by the instrument controls. Thehumidity was controlled by mixing streams of dry and wet nitrogen, witha total flow rate of 200 ml.min⁻¹ The relative humidity was measured bya calibrated Rotronic probe (dynamic range of 1.0-100% RH), located nearthe sample. The weight change, (mass relaxation) of the sample as afunction of % RH was constantly monitored by the microbalance (accuracy±0.005 mg).

Typically 5-20 mg of sample was placed in a tared mesh stainless steelbasket under ambient conditions. The sample was loaded and unloaded at40% RH and 25° C. (typical room conditions). A moisture sorptionisotherm was performed as outlined below (2 scans giving 1 completecycle). The standard isotherm was performed at 25° C. at 10% RHintervals over a 0.5-90% RH range.

TABLE 5 Method Parameters for SMS DVS Intrinsic Experiments ParametersValues Adsorption - Scan 1 40-90 Desorption/Adsorption - Scan 2 90-0,0-40 Intervals (% RH) 10 Number of Scans 4 Flow rate (ml · min⁻¹) 200Temperature (° C.) 25 Stability (° C.min⁻¹) 0.2 Sorption Time (hours) 6hour time out

The sample was recovered after completion of the isotherm andre-analyzed by XRPD.

Form A

Form A solvates were stable at 40° C. and 75% RH for at least a week.

Form B

The GVS isotherms of Form B at 25° C. showed that the uptake of water byForm B at 90% RH was less than 0.2%; therefore, Form B was nothygroscopic. No change in the XRPD pattern of the material after GVSanalysis was observed suggesting that Form B was stable under the GVSconditions.

No difference in the XRPD patterns of Form B before and after storage at25° C. and 92% RH for 12 days was observed suggesting that Form B wasstable under these conditions.

Form B was stable at 40° C. and 75% RH for at least a week.

Form C

Form C was stable at 40° C. and 75% RH for at least a week.

Form D

Form D was stable at 40° C. and 75% RH for at least a week.

Form E

Under the GVS conditions, Form E transformed to Form A.

A sample of Form E was laid on a glass slide then placed in a boxmaintained at 92% RH/25° C. Under these conditions, after a week Form Etransformed to Form A and a small amount of Form B.

Form E transformed to Form A at 40° C. and 75% RH within a week.

Form F

Form F transformed to Form A at 40° C. and 75% RH within a week.

Form G

Form G was stable at 40° C. and 75% RH for at least a week.

Form H

Form H was stable at 40° C. and 75% RH for at least a week.

Form I

A sample of Form I was laid on a glass slide then placed in a boxmaintained at 92% RH/25° C. Under these conditions, after a week Form Itransformed to Form B.

Form J

A sample of Form J was laid on a glass slide then placed in a boxmaintained at 92% RH/25° C. Under these conditions, after a week Form Jtransformed to Form B.

Example 7: Water Determination by Karl Fischer Titration (KF)

The water content of each sample was measured on a Mettler Toledo DL39Coulometer using Hydranal Coulomat AG reagent and an argon purge.Weighed solid samples were introduced into the vessel on a platinum TGApan which was connected to a subaseal to avoid water ingress.Approximately 10 mg of sample was used per titration and duplicatedeterminations were made.

In some embodiments, the water content for Form A was observed to be2.5% (w/w).

In some embodiments, the water content for Form C was observed to be0.4% (w/w).

In some embodiments, the water content for Form D was observed to be0.3% (w/w).

In some embodiments, the water content for Form J was observed to be0.3% (w/w).

Example 8: Thermodynamic Aqueous Solubility

Aqueous solubility was determined by suspending sufficient compound inwater to give a maximum final concentration of ≥20 mg.ml⁻¹ of the parentfree-form of the compound. The suspension was equilibrated at 25° C. for24 hours then the pH was measured. The suspension was then filteredthrough a glass fibre C filter into a 96 well plate unless statedotherwise. The filtrate was then diluted by a factor of 101.Quantitation was by HPLC with reference to a standard solution ofapproximately 0.1 mg.ml⁻¹ in DMSO. Different volumes of the standard,diluted and undiluted sample solutions were injected. The solubility wascalculated using the peak areas determined by integration of the peakfound at the same retention time as the principal peak in the standardinjection.

TABLE 6 HPLC Method Parameters for Solubility Measurements Type ofmethod: Reverse phase with gradient elution Column: Phenomenex Luna, C18(2) 5 μm 50 × 4.6 mm Column Temperature (° C.): 25 Standard Injections(μl): 1, 2, 3, 5, 7, 10 Test Injections (μl): 1, 2, 3, 10, 20, 50Detection: 260, 80 Wavelength, Bandwidth (nm): Flow Rate  2 (ml ·min⁻¹): Phase A: 0.1% TFA in water Phase B: 0.085% TFA in acetonitrileTime (min) % Phase A % Phase B Timetable: 0.0 95 5 1.0 80 20 2.3 5 953.3 5 95 3.5 95 5 4.4 95 5

Analysis was performed on an Agilent HP1100 series system equipped witha diode array detector and using ChemStation software vB.02.01-SR1.

TABLE 7 Solubility results Form Aqueous Solubility (mg/mL) A 0.01 B0.004

Example 9: Chemical Purity Determination

Purity analysis was performed by HPLC on an Agilent HP1100 series systemequipped with a diode array detector and using ChemStation softwarevB.02.01-SR1.

TABLE 8 HPLC Method Parameters for Chemical Purity Determinations SamplePreparation: 0.5 mg/ml in acetonitrile:water 1:1 (unless otherwisestated) Column: Supelco Ascentis Express C18, 100 × 4.6 mm, 2.7 μmColumn Temperature (° C.): 25 Injection (μl): 5 (unless otherwisestated) Detection: 255, 90 Wavelength, Bandwidth (nm): Flow Rate (ml ·min−1):   2.0 Phase A: 0.1% TFA in water Phase B: 0.085% TFA inacetonitrile Time (min) % Phase A % Phase B Timetable: 0 95 5 6 5 95 6.295 5 8 95 5

Samples of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidewere found to be greater than 95% pure. In some embodiments, samples of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidewere found to be greater than 95% pure, greater than 96% pure, greaterthan 97% pure, greater than 98% pure, or greater than 99% pure.

Example 10: Pharmaceutical Composition

Capsule Formulation

In one embodiment, capsule formulations of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidefor administration to humans are prepared with the followingingredients:

TABLE 9 Components of Capsule Formulation Quantity per Quantity perComponent Function Size 4 Capsule Size 1 Capsule crystalline4-[7-(6-cyano-5- Active 5 to 100 mg 50 to 500 mgtrifluoromethylpyridin-3- yl)-8-oxo-6-thioxo-5,7- diazaspiro[3.4]oct-5-yl]-2-fluoro-N- methylbenzamide Hypromellose, USP Capsule 1 capsule 1capsule Shell

The process to prepare crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein a capsule is as follows: Weigh the required amount of crystalline4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide,add into the appropriate size capsule, and close capsule.

In some embodiments, the capsules are stored at 25° C. for up to 48hours.

The examples and embodiments described herein are illustrative andvarious modifications or changes suggested to persons skilled in the artare to be included within this disclosure. As will be appreciated bythose skilled in the art, the specific components listed in the aboveexamples may be replaced with other functionally equivalent components,e.g., diluents, binders, lubricants, fillers, and the like.

What is claimed is:
 1. A crystalline Form D of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidethat is characterized as exhibiting at least one of: (a) an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 6.3±0.1°2-Theta, 13.9±0.1° 2-Theta, 16.4±0.1° 2-Theta, 17.0±0.1° 2-Theta,23.7±0.1° 2-Theta, and 24.8±0.1° 2-Theta; or (b) an X-Ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG.
 4. 2.The crystalline Form D of claim 1 that exhibits an X-ray powderdiffraction (XRPD) pattern with characteristic peaks at 6.3±0.1°2-Theta, 13.9±0.1° 2-Theta, 16.4±0.1° 2-Theta, 17.0±0.1° 2-Theta,23.7±0.1° 2-Theta, and 24.8±0.1° 2-Theta.
 3. The crystalline Form D ofclaim 1 that is further characterized as exhibiting at least one of: (a)a DSC thermogram substantially similar to the one set forth in FIG. 13;(b) a thermo-gravimetric analysis (TGA) thermogram substantially similarto the one set forth in FIG. 13; (c) a DSC thermogram with a firstendotherm having an onset temperature at about 122° C. and secondendotherm having an onset temperature at about 192° C.; or (d)substantially the same X-ray powder diffraction (XRPD) pattern poststorage at 40° C. and 75% RH for at least a week.
 4. The crystallineForm D of claim 1, further exhibiting a DSC thermogram with a firstendotherm having an onset temperature at about 122° C.
 5. Thecrystalline Form D of claim 3, wherein the crystalline form is furthercharacterized as having properties (a), (b), (c), and (d).
 6. Thecrystalline Form D of claim 1, wherein the crystalline form is atert-butyl methyl ether (TBME) solvate.
 7. A pharmaceutical compositioncomprising4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideand at least one additional ingredient that is a pharmaceuticallyacceptable carrier, diluent, or excipient, in which the4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamidein the composition comprises the crystalline Form D according toclaim
 1. 8. The pharmaceutical composition according to claim 7, whereinthe pharmaceutical composition is in a form formulated for oraladministration to a mammal.
 9. The pharmaceutical composition of claim8, wherein the mammal is a human.
 10. The pharmaceutical compositionaccording to claim 7, wherein the pharmaceutical composition is in anoral solid dosage form.
 11. The pharmaceutical composition according toclaim 9, wherein the pharmaceutical composition comprises a unit dosageform containing about 0.5 mg to about 1000 mg of the crystalline Form Dof4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamide.12. A method of treating prostate cancer in a mammal comprisingadministering the crystalline Form D of4-[7-(6-cyano-5-trifluoromethylpyridin-3-yl)-8-oxo-6-thioxo-5,7-diazaspiro[3.4]oct-5-yl]-2-fluoro-N-methylbenzamideaccording to claim 1 to the mammal in need of such treatment.
 13. Themethod of claim 12, wherein the prostate cancer is hormone sensitiveprostate cancer or hormone refractory prostate cancer.
 14. The method ofclaim 12, wherein the mammal is a human.
 15. The method of claim 13,wherein the mammal is a human.
 16. A method of treating prostate cancerin a mammal comprising administering the pharmaceutical compositionaccording to claim 7 to the mammal in need of such treatment.
 17. Themethod of claim 16, wherein the prostate cancer is hormone sensitiveprostate cancer or hormone refractory prostate cancer.
 18. The method ofclaim 16, wherein the mammal is a human.
 19. The method of claim 17,wherein the mammal is a human.